MN gene and protein

Abstract

A new gene-MN-and proteins/polypeptides encoded therefrom are disclosed. Recombinant nucleic acid molecules for expressing MN proteins/polypeptides and fusion proteins are provided. Expression of the MN gene is disclosed as being associated with tumorigenicity, and the invention concerns methods and compositions for detecting and/or quantitating MN antigen and/or MN-specific antibodies in vertebrate samples that are diagnostic/prognostic for neoplastic and pre-neoplastic disease. Test kits embodying the immunoassays of this invention are provided. MN-specific antibodies are disclosed that can be used diagnostically/prognostically, therapeutically, for imaging, and for affinity purification of MN proteins/polypeptides. Also provided are nucleic acid probes for the MN gene as well as test kits comprising said probes. The invention also concerns vaccines comprising MN proteins/polypeptides which are effective to immunize a vertebrate against neoplastic diseases associated with the expression of MN proteins. The invention still further concerns antisense nucleic acid sequences that can be used to inhibit MN gene expression.

What we claim is: 
     
       1. An isolated nucleic acid encoding a MN protein wherein the nucleotide sequence for said nucleic acid is selected from the group consisting of: (a) SEQ. ID. NO.: 1;   (b) nucleotide sequences that hybridize under stringent conditions to SEQ. ID. No.: 1 or to its complementary strand; and   (c) nucleotide sequences that differ from SEQ. ID. NO.: 1 and from the nucleotide sequences of (b) in codon sequence due to the degeneracy of the genetic code.   
     
     
       2. The isolated nucleic acid according to claim 1 wherein the selected nucleotide sequence for said isolated nucleic acid is SEQ. ID. NO.: 1. 
     
     
       3. The isolated nucleic acid according to claim 1 wherein the selected nucleotide sequence is contained in a vector. 
     
     
       4. The isolated nucleic acid according to claim 2 wherein SEQ. ID. NO.: 1 is contained in a vector. 
     
     
       5. The isolated nucleic acid according to claim 3 wherein said vector is a bacterial cloning vector. 
     
     
       6. The isolated nucleic acid according to claim 5 wherein said vector is pGEX-3X. 
     
     
       7. The isolated nucleic acid according to claim 1 wherein fragments of said isolated nucleic acid are nucleic acid probes which specifically hybridize under stringent conditions to nucleic acid sequences encoding MN proteins or to the complementary sequences to those encoding MN proteins and do not hybridize under stringent conditions to nucleic acid sequences encoding carbonic anhydrase proteins or to sequences complementary to those encoding carbonic anhydrase proteins. 
     
     
       8. The isolated nucleic acid according to claim 1 wherein fragments of said nucleic acid are polymerase chain reaction primers for segments of MN genes wherein said primers specifically hybridize under stringent conditions to nucleic acid sequences encoding MN proteins or to sequences complementary to those encoding MN proteins, but do not hybridize under stringent conditions to nucleic acid sequences encoding carbonic anhydrase proteins or to sequences complementary to those encoding carbonic anhydrase proteins. 
     
     
       9. The isolated nucleic acid according to claim 1 wherein said nucleic acid is DNA. 
     
     
       10. The isolated nucleic acid according to claim 3 wherein said vector is a cloning vector comprising a first and a second restriction endonuclease recognition site, and said nucleic acid is inserted between said first and second restriction sites. 
     
     
       11. The isolated nucleic acid according to claim 10 wherein said nucleic acid is operatively linked to an expression control sequence in said vector. 
     
     
       12. A unicellular host which is either prokaryotic or eukaryotic that is transformed or transfected with the isolated nucleic acid operatively linked to an expression control sequence in a vector according to claim 11. 
     
     
       13. A nucleic acid probe which is selected from the group consisting of: a. SEQ. ID. NO.: 1; and   b. nucleotide sequences which hybridize under stringent conditions to SEQ. ID. NO.: 1 or to the complementary strand to SEQ. ID. NO.: 1, wherein said nucleotide sequences do not hybridize under stringent conditions to nucleic acid sequences encoding carbonic anhydrase proteins or to sequences complementary to those encoding carbonic anhydrase proteins.   
     
     
       14. The nucleic acid probe according to claim 13 which is SEQ. ID. NO. 1. 
     
     
       15. A test kit comprising the nucleic acid probe according to claim 13 and a means to enable the visualization of said nucleic acid probe once hybridized to an appropriate MN gene target. 
     
     
       16. An isolated nucleic acid encoding a MN protein to which monoclonal antibodies designated M75 produced by the hybridoma VU-M75 deposited at the American Type Culture Collection (ATCC) in Rockville, Md. in the United States of America under ATCC No. HB 11128, specifically bind, wherein the nucleotide sequence for said nucleic acid is selected from the group consisting of: (a) SEQ. ID. NO.: 1;   (b) nucleotide sequences that hybridize under stringent conditions to SEQ. ID. NO.: 1 or to the complementary strand to SEQ. ID. NO.: 1; and   (c) nucleotide sequences that differ from SEQ. ID. NO.: 1 and from the nucleotide sequences of (b) in codon sequence due to the degeneracy of the genetic code.   
     
     
       17. An isolated nucleic acid encoding a fusion protein consisting essentially of a MN protein and a non-MN protein wherein the nucleotide sequence for the portion of the nucleic acid encoding the MN protein is selected from the group consisting of: (a) SEQ. ID. NO.: 1;   (b) nucleotide sequences that hybridize under stringent conditions to SEQ. ID. NO.: 1 or to the complementary strand to SEQ. ID. NO.: 1; and   (c) nucleotide sequences that differ from SEQ. ID. NO.: 1 and from the nucleotide sequences of (b) in codon sequence due to the degeneracy of the genetic code; and wherein the non-MN protein is not immunogenic.     
     
     
       18. The isolated nucleic acid according to claim 17 wherein said isolated nucleic acid sequence is contained in a vector. 
     
     
       19. The isolated nucleic acid according to claim 18 wherein said vector is pGEX-3X. 
     
     
       20. The isolated nucleic acid according to claim 17 wherein said non-MN protein is the alpha-peptide region of beta-galactosidase or the carboxyl terminus of glutathione S-transferase.